Quantitative receptor autoradiography using [3H]Ultrofilm: application to multiple benzodiazepine receptors

Unnerstall, James R.; Niehoff, Debra L.; Kuhar, Michael J.; Palacios, J.M.

doi:10.1016/0165-0270(82)90016-4

Cited by 521 publications

(95 citation statements)

References 23 publications

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“…The response of the film to radioactive exposure was found to be similar to that reported by other authors (Unnerstall et al 1982;Kuhar and Unnerstall 1985; (A) Aliquots of brain pastes were mixed with increasing quantities of radioactivity and frozen. Representative tissue paste sections were cut in a cryostat and collected, and their radioactivities were determined.…”

Section: Discussionsupporting

confidence: 81%

“…Davenport et al 1988;Ito et al 1995). For approximation, interpolation and calibration, in contrast with previous studies (Sokoloff et al 1977;Unnerstall et al 1982;Pan et al 1983;Miller et al 1988), a novel mathematical approach was introduced. The exponential model developed in this study [ROD ϭ p 1 (1 Ϫ exp[Ϫp 2 x])] utilizes two parameters and describes the ROD vs radioactivity relationship appropriately (r 2 Ն0.999) between 0 and 240 (out of 255) GVs with Hyperfilm-␤max.…”

Section: Discussionmentioning

confidence: 99%

“…The two currently used radioactive standard scales are the tissue pasteand plastic-based ones. Tissue paste standards are prepared by the investigator by mixing increasing amounts of the radioisotope to be quantified with tissue paste (Unnerstall et al 1982;Davenport et al 1988). Because the paste is derived from the tissue under investigation, the quenching of radiation due to variations in the media and the thickness and mounting of the specimens is essentially the same in both the standards and the tissue sections.…”

mentioning

confidence: 99%

“…In the absence of an explicit theory describing the complex relationship between OD and radioactivity, third-order (Sokoloff et al 1977;Goochee et al 1980) and fourth-order (Pan et al 1983;Baskin and Stahl 1993) polynomial, power (Unnerstall et al 1982), or logistic functions (Miller et al 1988) have been utilized for approximation and interpolation of the autoradiographic data of the standard scales. The application of the above-mentioned functions is based merely on empirical observations, with the intention of finding the best fit (Baskin and Wimpy 1989).…”

mentioning

confidence: 99%

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A New Quantitative Film Autoradiographic Method of Quantifying mRNA Transcripts for In Situ Hybridization

Palfi

Hatvani

Gulya

1998

J Histochem Cytochem.

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SUMMARYWe developed and tested a novel quantitative method for the quantification of film autoradiographs, involving a mathematical model and a dot-blot-based membrane standard scale. The exponential model introduced here, ROD ϭ p 1 (1 Ϫ exp[p 2 x]), appropriately (r 2 Ͼ0.999), describes the relation between relative optical density (ROD) and radioactivity (x) in the range between 0 and 240 gray scale values (using a 256-gray scale level digitizer). By means of this model, standard curves with distinct quenching properties can be exactly interconverted, permitting the tissue-equivalent calibration of different standard scales. The membrane standard scale employed here has several advantages, including the flexible radioactivity range, the facile and rapid preparation technique, and the compact size. The feasibility of the quantification procedure is exemplified by the comparative quantification of multiple calmodulin mRNAs in the rat brain by in situ hybridization with [ 35 S]-cRNA probes. The procedure for quantification provides a significant improvement in that the direct and exact comparison of radiolabeled species, even from different experiments, can be reliably performed. Further, the procedure can be adapted to the quantification of autoradiographs produced by other methods. (J Histochem Cytochem 46:1141-1149, 1998)

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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A New Quantitative Film Autoradiographic Method of Quantifying mRNA Transcripts for In Situ Hybridization

Palfi

Hatvani

Gulya

1998

J Histochem Cytochem.

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“…The films were analyzed densitometrically on an Apple computer-based system (Baskin et al, 1986), and regions of interest within the hippocampus were referenced to tritiated brain mash standards prepared as previously described (Unnerstall et al, 1982). Optical densities over the regions of interest were converted to pmole/mg protein according to the conversion of Unnerstall et al (1982), and FLU receptor number and affinity were determined by Scatchard analysis. Two hippocampal regions were chosen for analysis: stratum radiatum ofCAl and the dentate gyrus molecular layer.…”

Section: Methodsmentioning

confidence: 99%

Inhibition in kainate-lesioned hyperexcitable hippocampi: physiologic, autoradiographic, and immunocytochemical observations

et al. 1988

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Following kainate lesions of hippocampal subfield CA3, the remaining CA 1 pyramidal cells become hyperexcitable. This lesion is of interest because, morphologically, it resembles the damage often seen in cases of temporal lobe epilepsy; it may provide insight into the consequences of such cell loss in humans. The hyperexcitability in CA 1 is associated with a loss of both early and late IPSPs. At long postlesion latencies (2–4 months) inhibition shows partial recovery and the hyperexcitability subsides. The intent of the present work was to determine if alterations in CA 1 excitability and functional inhibition postlesion are correlated with changes in morphologic and physiologic indicators of inhibitory interneuron function or with alterations in binding sites for inhibitory transmitters. Using GAD immunocytochemistry, we found no acute or chronic lesion-induced decrease in numbers of CA 1 interneurons or in qualitative characteristics of the pericellular distribution of their terminals in CA 1 stratum pyramidale. Intracellular recordings from identified cells in CA 1 indicated that putative interneurons were viable in hyperexcitable tissue. It was further observed that “recovery” in tissue studied 2–4 months postlesion primarily involved the early IPSP; the late IPSP failed to reappear. Quantitative in vitro autoradiographic analysis of 3H-flunitrazepam--a marker for the early IPSP associated GABAA receptor complex--indicated that hyperexcitability was associated with an increase in GABAA receptor number in CA 1; receptor binding returned to normal at long postlesion latencies as the early IPSP returned and hyperexcitability subsided. Finally, hyperexcitable pyramidal cells were found to retain their responsivity to exogenously applied GABA. These data indicate that much of the cellular machinery necessary for inhibition is retained in CA 1, despite lesion-induced hyperexcitability. We suggest that the acute loss of the IPSP after kainate lesion is due to a transient disconnection between inhibitory and excitatory elements in CA 1 and/or to a loss of normal afferent drive from CA3 onto some CA 1 interneurons. We further suggest that incomplete recovery can be explained by abnormalities that occur as neuroplastic rearrangements in response to deafferentation of CA 1. The relevance of these studies to human hippocampal necrosis and to other models of focal epilepsy is discussed.

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Autoradiographic localization of neurotransmitter binding sites in the hypoglossal and motor trigeminal nuclei of the rat

Manaker

Zucchi

1998

Synapse

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The hypoglossal and motor trigeminal nuclei contain somatic motoneurons innervating the tongue, jaw, and palate. These two cranial motor nuclei are myotopically organized and contain neurotransmitter binding sites for thyrotropin-releasing hormone, substance P, and serotonin. Quantitative autoradiography was used to localize thyrotropin-releasing hormone, substance P, and serotonin-1A and serotonin-1B binding sites in the hypoglossal and motor trigeminal nuclei and to relate the relative distributions of these binding sites to the myotopic organizations of the two nuclei. In the hypoglossal nucleus, high-to-moderate concentrations of all four binding sites were present in the dorsal and ventromedial subnuclei, whereas low concentrations were noted in the ventrolateral subnucleus. In the motor trigeminal nucleus, high concentrations of serotonin-1B, moderate densities of thyrotropin-releasing hormone, and low levels of substance P and serotonin-1A binding sites were present in both the ventromedial and dorsolateral subnuclei. These observations demonstrate that neurotransmitter binding sites in the hypoglossal and motor trigeminal nuclei are heterogeneously localized and that their distributions correspond to the previously described myotopic organizations of each nucleus.

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Quantitative receptor autoradiography using [3H]Ultrofilm: application to multiple benzodiazepine receptors

Cited by 521 publications

References 23 publications

A New Quantitative Film Autoradiographic Method of Quantifying mRNA Transcripts for In Situ Hybridization

A New Quantitative Film Autoradiographic Method of Quantifying mRNA Transcripts for In Situ Hybridization

Inhibition in kainate-lesioned hyperexcitable hippocampi: physiologic, autoradiographic, and immunocytochemical observations

Autoradiographic localization of neurotransmitter binding sites in the hypoglossal and motor trigeminal nuclei of the rat

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